Rapid development of naked malting barley germplasm through targeted mutagenesis.

IF 2.6 3区 农林科学 Q1 AGRONOMY Molecular Breeding Pub Date : 2025-03-07 eCollection Date: 2025-03-01 DOI:10.1007/s11032-025-01553-5
Hiroshi Hisano, Hiroaki Sakai, Mika Hamaoka, Hiromi Munemori, Fumitaka Abe, Brigid Meints, Kazuhiro Sato, Patrick M Hayes
{"title":"Rapid development of naked malting barley germplasm through targeted mutagenesis.","authors":"Hiroshi Hisano, Hiroaki Sakai, Mika Hamaoka, Hiromi Munemori, Fumitaka Abe, Brigid Meints, Kazuhiro Sato, Patrick M Hayes","doi":"10.1007/s11032-025-01553-5","DOIUrl":null,"url":null,"abstract":"<p><p>Covered barley (<i>Hordeum vulgare</i>) has historically been preferred for malting, as the husk in this plant protects the embryo during harvest and acts as a filter during brewing. Naked barley, which is typically used as food, has the potential to be used in brewing due to recent technical advances, but the grains contain higher levels of β-glucan and polyphenols, which are undesirable in brewing. Introducing the naked trait into brewing cultivars through crossing is time-consuming due to the need to eliminate these undesirable traits. In this study, we rapidly developed naked barley that is potentially suitable for malting by introducing targeted mutations into <i>Nudum</i> (<i>NUD</i>) using CRISPR/Cas9-mediated targeted mutagenesis. The doubled haploid line 'DH120366', which was used as the parental line, was derived from a cross between two covered malting barley cultivars. We generated CRISPR/Cas9-mediated targeted mutagenized barley harboring mutations in <i>NUD</i> via <i>Agrobacterium tumefaciens</i>-mediated transformation and confirmed the presence of mosaic mutations in one individual from among 16 T<sub>0</sub> transformants. We sowed T<sub>1</sub> grains exhibiting the naked trait and sequenced the <i>NUD</i> gene in these T<sub>1</sub> seedlings, identifying two types of mutations. Shotgun high-throughput whole-genome sequencing confirmed the absence of the transgene in at least one <i>nud</i> mutant line following <i>k</i>-mer-based analysis. Cultivation in a closed growth chamber revealed no significant differences in agronomic traits between the <i>nud</i> mutants and the wild type. This study demonstrates the feasibility of rapidly developing naked barley with potential use for malting and brewing by targeting only <i>NUD</i> via targeted mutagenesis.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s11032-025-01553-5.</p>","PeriodicalId":18769,"journal":{"name":"Molecular Breeding","volume":"45 3","pages":"32"},"PeriodicalIF":2.6000,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11889295/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Breeding","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1007/s11032-025-01553-5","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/3/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"AGRONOMY","Score":null,"Total":0}
引用次数: 0

Abstract

Covered barley (Hordeum vulgare) has historically been preferred for malting, as the husk in this plant protects the embryo during harvest and acts as a filter during brewing. Naked barley, which is typically used as food, has the potential to be used in brewing due to recent technical advances, but the grains contain higher levels of β-glucan and polyphenols, which are undesirable in brewing. Introducing the naked trait into brewing cultivars through crossing is time-consuming due to the need to eliminate these undesirable traits. In this study, we rapidly developed naked barley that is potentially suitable for malting by introducing targeted mutations into Nudum (NUD) using CRISPR/Cas9-mediated targeted mutagenesis. The doubled haploid line 'DH120366', which was used as the parental line, was derived from a cross between two covered malting barley cultivars. We generated CRISPR/Cas9-mediated targeted mutagenized barley harboring mutations in NUD via Agrobacterium tumefaciens-mediated transformation and confirmed the presence of mosaic mutations in one individual from among 16 T0 transformants. We sowed T1 grains exhibiting the naked trait and sequenced the NUD gene in these T1 seedlings, identifying two types of mutations. Shotgun high-throughput whole-genome sequencing confirmed the absence of the transgene in at least one nud mutant line following k-mer-based analysis. Cultivation in a closed growth chamber revealed no significant differences in agronomic traits between the nud mutants and the wild type. This study demonstrates the feasibility of rapidly developing naked barley with potential use for malting and brewing by targeting only NUD via targeted mutagenesis.

Supplementary information: The online version contains supplementary material available at 10.1007/s11032-025-01553-5.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
求助全文
约1分钟内获得全文 去求助
来源期刊
Molecular Breeding
Molecular Breeding 农林科学-农艺学
CiteScore
5.60
自引率
6.50%
发文量
67
审稿时长
1.5 months
期刊介绍: Molecular Breeding is an international journal publishing papers on applications of plant molecular biology, i.e., research most likely leading to practical applications. The practical applications might relate to the Developing as well as the industrialised World and have demonstrable benefits for the seed industry, farmers, processing industry, the environment and the consumer. All papers published should contribute to the understanding and progress of modern plant breeding, encompassing the scientific disciplines of molecular biology, biochemistry, genetics, physiology, pathology, plant breeding, and ecology among others. Molecular Breeding welcomes the following categories of papers: full papers, short communications, papers describing novel methods and review papers. All submission will be subject to peer review ensuring the highest possible scientific quality standards. Molecular Breeding core areas: Molecular Breeding will consider manuscripts describing contemporary methods of molecular genetics and genomic analysis, structural and functional genomics in crops, proteomics and metabolic profiling, abiotic stress and field evaluation of transgenic crops containing particular traits. Manuscripts on marker assisted breeding are also of major interest, in particular novel approaches and new results of marker assisted breeding, QTL cloning, integration of conventional and marker assisted breeding, and QTL studies in crop plants.
期刊最新文献
Using genomic selection to correct pedigree errors in kiwiberry breeding. Genomic selection in a kiwiberry breeding programme: integrating intra- and inter-specific crossing. Rapid development of naked malting barley germplasm through targeted mutagenesis. CsKIP1.7A, a gene involved in fruit development, contributes to the yield heterosis formation of hybrid F1 in cucumber. A novel trait to reduce the mechanical damage of peach fruits at harvest: The first genetic dissection study for peduncle length.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1